Abstract
Metabolically competent, inexpensive, and robust in vitro cell models are needed for studying liver drug-metabolizing enzymes and hepatotoxicity. Human hepatoma HuH-7 cells develop into a differentiated in vitro model resembling primary human hepatocytes after a 2-week dimethyl sulfoxide (DMSO) treatment. DMSO-treated HuH-7 cells express elevated cytochrome P450 3A4 (CYP3A4) enzyme gene expression and activity compared to untreated HuH-7 cells. This cell model could be used to study CYP3A4 inhibition by reversible and time-dependent inhibitors, including drugs, food-related substances, and environmental chemicals. The DMSO-treated HuH-7 model is also a suitable tool for investigating hepatotoxicity. This chapter describes a detailed methodology for developing DMSO-treated HuH-7 cells, which are subsequently used for CYP3A4 inhibition and hepatotoxicity studies.
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References
Lin J, Schyschka L, Muhl-Benninghaus R, Neumann J, Hao L, Nussler N, Dooley S, Liu L, Stockle U, Nussler AK, Ehnert S (2012) Comparative analysis of phase I and II enzyme activities in 5 hepatic cell lines identifies Huh-7 and HCC-T cells with the highest potential to study drug metabolism. Arch Toxicol 86(1):87–95. doi:10.1007/s00204-011-0733-y
Donato MT, Jover R, Gomez-Lechon MJ (2013) Hepatic cell lines for drug hepatotoxicity testing: limitations and strategies to upgrade their metabolic competence by gene engineering. Curr Drug Metab 14(9):946–968
Nakabayashi H, Taketa K, Miyano K, Yamane T, Sato J (1982) Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. Cancer Res 42(9):3858–3863
Choi S, Sainz B Jr, Corcoran P, Uprichard S, Jeong H (2009) Characterization of increased drug metabolism activity in dimethyl sulfoxide (DMSO)-treated Huh7 hepatoma cells. Xenobiotica 39(3):205–217. doi:10.1080/00498250802613620
Liu Y, Flynn TJ, Xia M, Wiesenfeld PL, Ferguson MS (2015) Evaluation of CYP3A4 inhibition and hepatotoxicity using DMSO-treated human hepatoma HuH-7 cells. Cell Biol Toxicol 31(4-5):221–230. doi:10.1007/s10565-015-9306-9
Hisaka A, Ohno Y, Yamamoto T, Suzuki H (2010) Prediction of pharmacokinetic drug-drug interaction caused by changes in cytochrome P450 activity using in vivo information. Pharmacol Ther 125(2):230–248. doi:10.1016/j.pharmthera.2009.10.011
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Liu, Y. (2016). Study Liver Cytochrome P450 3A4 Inhibition and Hepatotoxicity Using DMSO-Differentiated HuH-7 Cells. In: Zhu, H., Xia, M. (eds) High-Throughput Screening Assays in Toxicology. Methods in Molecular Biology, vol 1473. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6346-1_7
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DOI: https://doi.org/10.1007/978-1-4939-6346-1_7
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6344-7
Online ISBN: 978-1-4939-6346-1
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